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Dried blood spot screening of lysosomal acid lipase deficiency and confirmatory studies in Spanish suspected patients
Abstracts / Molecular Genetics and Metabolism 117 (2016) S14–S124
cholesterol, TG concentration, ALT and AST activity were assessed in both sets. Plasma ChT activity was evaluated by fluorimeter assay and CCL18/PARC concentration by ELISA. Leukocyte LAL activity was measured using fluorescence based enzyme assay with slightly modifications on previously published. Patients with anomalous lipid/ liver levels and/or biomarkers, underwent LIPA gene sequenciation. Eight subjects from Set 1 and twenty from Set 2 showed altered lipid/ liver profile and/or biomarkers and/or null/reduced LAL activity. LIPA gene sequencing identified in Set 2, one subject carried a novel mutation in promoter region, one homozygous for the most prevalent mutation [p.Q298 = (E8SJM)] and one complex heterozygous for E8SJM and novel nonsense mutation. In Set 1 we had not identified any mutation. According to our results, the analysis of plasma ChT activity and CCL18/ PARC concentration, combined with lipid/liver profile is an useful approach to identify LALD patients who should undergo testing for enzymatic/genetic diagnostic. doi:10.1016/j.ymgme.2015.12.215
58 Dried blood spot screening of lysosomal acid lipase deficiency and confirmatory studies in Spanish suspected patients Jorge J. Cebollaa,b,c,d,e, Pilar Iruna,b,c,f, Luisa Gonzalez-Dieguezg, Pablo del Valle Loarteh, Miguel Angel Barba-Romeroi, Inmaculada GarciaJimeneza, Ignacio Ros Arnala, Pilar Giraldoa,b,c,e,f, aMiguel Servet University Hospital, Zaragoza, Spain, bAragón Health Sciencies Institute (I+CS), Zaragoza, Spain, cAragón Health Research Institute (IIS Aragón), Zaragoza, Spain, dUniversidad de Zaragoza, Zaragoza, Spain, eSpanish Foundation of Gaucher Disease and other Lysosomal Disorders (FEETEG), Zaragoza, Spain, fCentro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER, ISCIII), Zaragoza, Spain, gAsturias Central University Hospital, Oviedo, Spain, hSevero Ochoa University Hospital, Leganés, Spain, iAlbacete General University Hospital, Albacete, Spain Lysosomal acid lipase deficiency (LALD) is an inborn error of metabolism, triggering accumulation of cholesterol esterified and/or triglycerides in lysosomes. The main manifestations are dyslipidemia, liver alterations, cardiovascular disease and gastrointestinal disturbances. LALD screening is performed by easy-handle procedure from dried blood spots (DBS), however confirmatory studies are mandatory to assess LALD status. The purposes were: Firstly, LAL DBS screening in clinically compatible probands and identification of potentially LALD patients. Secondly, confirm LALD status through characterization of biomarkers and LIPA gene mutations. Lastly, accomplish family studies. For that, probands were recruited according to clinical findings. DBS were obtained and LAL activity assessed in all probands using a fluorescence based enzyme assay with slightly modifications on previously published; Null/almost null LAL activity were considered LALD potentially patients and required deeper studies in whole blood sample. LALD status was confirmed by determining plasma biomarkers (Chitotriosidase activity, CCL18/PARC concentration and 7-Ketocholesterol concentration) and by LIPA gene sequencing. 65 subjects (23 females and 42 males) were referred: 38 pediatrics, nonoverweighed and hepatomegaly (44.7%) and low cholesterol-HDL levels (37.1%) are the most common findings; 25 adults, non-overweighed, splenomegaly and steatosis (32.0%) and low cholesterol-HDL levels (56.0%) are prevalent. LAL DBS [median(IQR) nmol/punch/h] in pediatrics were 0.69(0.54-1.15) whereas 0.66(0.31-0.97) in adults; 3 pediatrics and 4 adults showed DBS activity under 0.03 nmol/punch/h, therefore were assessed biomarkers: 5 of 7 patients presented higher levels than intralaboratory cut-off values. LIPA sequencing identified 3 homozygous for most prevalent mutation p.Q298 = (E8SJM), 3 homozygous for p.H129R and one complex heterozygous for E8SJM and novel nonsense
S33
mutation. Family studies identified 9 carriers and 1 affected more. These results support the efficiency of this screening approach, identifying eight LALD patients without major extraction and shipping requirements. Furthermore, confirmatory studies assess mutations and biomarkers levels in those, to well-characterization and orientate family studies. doi:10.1016/j.ymgme.2015.12.216
59 Adhesion to the enzymatic replacement therapy in Gaucher disease and the effect on the bone health Magdalena Cerón, Hospital Infantil de Mexico Federico Gómez, Mexico City, Mexico Gaucher disease (GD) is a lysosomal disorder with high prevalence of bone damage (75%) bone crisis (BC), growth delay and characteristic changes in magnetic resonance image (MRI). Enzymatic replacement therapy (ERT) has as its goal to control BC, bone lesion and avoiding the development of skeletal abnormalities and as a result the improvement of the quality of life. Objective: To describe the impact of adherence to ERT on the bone health of 4 patients with GD in a tertiary level hospital. Methods: Four patients with GD under ERT, 3 with imiglucerase, 1 with velaglucerase, for 3 years at Hospital Infantil de México Federico Gómez. MRI of femur at the beginning and at later control. Results: Initial assessment (IA)
Following assessment (FA)
Patient 1: Adherence 91%, Growth/height to age (G/H) 89.1% to 98.3%, flask deformity, infiltration following decrease. Patient 2: Adherence 95%, G/H age 95% to 100%, flask deformity, initial infiltration and following decrease, cortical thinning and then cortical thickening Patient 3: Adherence 100%,G/H 87.5% to 93%, initial infiltration, pending following assestment (PFA) Patient 4: Adherence 100%, G/H both 100%, flask deformity, PFA
Discussion: The study evaluated the presence of bone complication at the beginning and throughout the treatment, as well as femur MRI. Adherence to the treatment was between 90% and 100%. The evolution was good without BC and recovery growth rate and improvement in MRI. Conclusion: Strict adherence to the treatment is related to an improvement in bone health and impact in a better quality of life. doi:10.1016/j.ymgme.2015.12.217
60 Differential response of glomerular parietal epithelial cells and podocytes to enzyme replacement therapy in Fabry nephropathy Fu-Pang Changa,b, Michael Mauerc, Alexey Sokolovskiya, Camilla Tøndeld,e, Einar Svarstadd,e, Behzad Najafiana, aUniversity of Washington, Seattle, WA, United States, bTaipei Veterans General Hospital, Taipei, Taiwan, cUniversity of Minnesota, Minneapolis, MN, United States, dUniversity of Bergen, Bergen, Norway, eHaukeland University Hospital, Bergen, Norway Podocyte injury is critical in progression of Fabry nephropathy (FN). Podocytes do not efficiently proliferate. Parietal epithelial cells (PEC) may replace lost podocytes, thus, PEC clearance from globotriaosylceramide (GL-3) may be important in podocyte regeneration in FN. Podocytes partially lose GL-3 after 1 year of ERT (JASN 25:175A, 2014). Here, we examined whether PEC lose GL-3 with ERT. Kidney biopsies at baseline (ERT-naive) and after 1 year of ERT (1 mg/kg/2 weeks agalsidase-B) from
cholesterol, TG concentration, ALT and AST activity were assessed in both sets. Plasma ChT activity was evaluated by fluorimeter assay and CCL18/PARC concentration by ELISA. Leukocyte LAL activity was measured using fluorescence based enzyme assay with slightly modifications on previously published. Patients with anomalous lipid/ liver levels and/or biomarkers, underwent LIPA gene sequenciation. Eight subjects from Set 1 and twenty from Set 2 showed altered lipid/ liver profile and/or biomarkers and/or null/reduced LAL activity. LIPA gene sequencing identified in Set 2, one subject carried a novel mutation in promoter region, one homozygous for the most prevalent mutation [p.Q298 = (E8SJM)] and one complex heterozygous for E8SJM and novel nonsense mutation. In Set 1 we had not identified any mutation. According to our results, the analysis of plasma ChT activity and CCL18/ PARC concentration, combined with lipid/liver profile is an useful approach to identify LALD patients who should undergo testing for enzymatic/genetic diagnostic. doi:10.1016/j.ymgme.2015.12.215
58 Dried blood spot screening of lysosomal acid lipase deficiency and confirmatory studies in Spanish suspected patients Jorge J. Cebollaa,b,c,d,e, Pilar Iruna,b,c,f, Luisa Gonzalez-Dieguezg, Pablo del Valle Loarteh, Miguel Angel Barba-Romeroi, Inmaculada GarciaJimeneza, Ignacio Ros Arnala, Pilar Giraldoa,b,c,e,f, aMiguel Servet University Hospital, Zaragoza, Spain, bAragón Health Sciencies Institute (I+CS), Zaragoza, Spain, cAragón Health Research Institute (IIS Aragón), Zaragoza, Spain, dUniversidad de Zaragoza, Zaragoza, Spain, eSpanish Foundation of Gaucher Disease and other Lysosomal Disorders (FEETEG), Zaragoza, Spain, fCentro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER, ISCIII), Zaragoza, Spain, gAsturias Central University Hospital, Oviedo, Spain, hSevero Ochoa University Hospital, Leganés, Spain, iAlbacete General University Hospital, Albacete, Spain Lysosomal acid lipase deficiency (LALD) is an inborn error of metabolism, triggering accumulation of cholesterol esterified and/or triglycerides in lysosomes. The main manifestations are dyslipidemia, liver alterations, cardiovascular disease and gastrointestinal disturbances. LALD screening is performed by easy-handle procedure from dried blood spots (DBS), however confirmatory studies are mandatory to assess LALD status. The purposes were: Firstly, LAL DBS screening in clinically compatible probands and identification of potentially LALD patients. Secondly, confirm LALD status through characterization of biomarkers and LIPA gene mutations. Lastly, accomplish family studies. For that, probands were recruited according to clinical findings. DBS were obtained and LAL activity assessed in all probands using a fluorescence based enzyme assay with slightly modifications on previously published; Null/almost null LAL activity were considered LALD potentially patients and required deeper studies in whole blood sample. LALD status was confirmed by determining plasma biomarkers (Chitotriosidase activity, CCL18/PARC concentration and 7-Ketocholesterol concentration) and by LIPA gene sequencing. 65 subjects (23 females and 42 males) were referred: 38 pediatrics, nonoverweighed and hepatomegaly (44.7%) and low cholesterol-HDL levels (37.1%) are the most common findings; 25 adults, non-overweighed, splenomegaly and steatosis (32.0%) and low cholesterol-HDL levels (56.0%) are prevalent. LAL DBS [median(IQR) nmol/punch/h] in pediatrics were 0.69(0.54-1.15) whereas 0.66(0.31-0.97) in adults; 3 pediatrics and 4 adults showed DBS activity under 0.03 nmol/punch/h, therefore were assessed biomarkers: 5 of 7 patients presented higher levels than intralaboratory cut-off values. LIPA sequencing identified 3 homozygous for most prevalent mutation p.Q298 = (E8SJM), 3 homozygous for p.H129R and one complex heterozygous for E8SJM and novel nonsense
S33
mutation. Family studies identified 9 carriers and 1 affected more. These results support the efficiency of this screening approach, identifying eight LALD patients without major extraction and shipping requirements. Furthermore, confirmatory studies assess mutations and biomarkers levels in those, to well-characterization and orientate family studies. doi:10.1016/j.ymgme.2015.12.216
59 Adhesion to the enzymatic replacement therapy in Gaucher disease and the effect on the bone health Magdalena Cerón, Hospital Infantil de Mexico Federico Gómez, Mexico City, Mexico Gaucher disease (GD) is a lysosomal disorder with high prevalence of bone damage (75%) bone crisis (BC), growth delay and characteristic changes in magnetic resonance image (MRI). Enzymatic replacement therapy (ERT) has as its goal to control BC, bone lesion and avoiding the development of skeletal abnormalities and as a result the improvement of the quality of life. Objective: To describe the impact of adherence to ERT on the bone health of 4 patients with GD in a tertiary level hospital. Methods: Four patients with GD under ERT, 3 with imiglucerase, 1 with velaglucerase, for 3 years at Hospital Infantil de México Federico Gómez. MRI of femur at the beginning and at later control. Results: Initial assessment (IA)
Following assessment (FA)
Patient 1: Adherence 91%, Growth/height to age (G/H) 89.1% to 98.3%, flask deformity, infiltration following decrease. Patient 2: Adherence 95%, G/H age 95% to 100%, flask deformity, initial infiltration and following decrease, cortical thinning and then cortical thickening Patient 3: Adherence 100%,G/H 87.5% to 93%, initial infiltration, pending following assestment (PFA) Patient 4: Adherence 100%, G/H both 100%, flask deformity, PFA
Discussion: The study evaluated the presence of bone complication at the beginning and throughout the treatment, as well as femur MRI. Adherence to the treatment was between 90% and 100%. The evolution was good without BC and recovery growth rate and improvement in MRI. Conclusion: Strict adherence to the treatment is related to an improvement in bone health and impact in a better quality of life. doi:10.1016/j.ymgme.2015.12.217
60 Differential response of glomerular parietal epithelial cells and podocytes to enzyme replacement therapy in Fabry nephropathy Fu-Pang Changa,b, Michael Mauerc, Alexey Sokolovskiya, Camilla Tøndeld,e, Einar Svarstadd,e, Behzad Najafiana, aUniversity of Washington, Seattle, WA, United States, bTaipei Veterans General Hospital, Taipei, Taiwan, cUniversity of Minnesota, Minneapolis, MN, United States, dUniversity of Bergen, Bergen, Norway, eHaukeland University Hospital, Bergen, Norway Podocyte injury is critical in progression of Fabry nephropathy (FN). Podocytes do not efficiently proliferate. Parietal epithelial cells (PEC) may replace lost podocytes, thus, PEC clearance from globotriaosylceramide (GL-3) may be important in podocyte regeneration in FN. Podocytes partially lose GL-3 after 1 year of ERT (JASN 25:175A, 2014). Here, we examined whether PEC lose GL-3 with ERT. Kidney biopsies at baseline (ERT-naive) and after 1 year of ERT (1 mg/kg/2 weeks agalsidase-B) from